The present invention relates to antibacterials; more specifically to mutilins. Pleuromutilins are naturally occurring antibiotics which have antimycoplasmal activity and modest antibacterial activity. We have found mutilins having the principal ring structure of naturally occurring pleuromutilins which have improved antimicrobial, e.g. antibacterial activity.
In one aspect the present invention provides a compound selected from 14xe2x80x94Oxe2x80x94[(cycloalkyl-sulfanyl)acetyl]mutilins; 14xe2x80x94Oxe2x80x94[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins; 14xe2x80x94Oxe2x80x94[(cycloalkoxy)acetyl] mutilins; and14xe2x80x94Oxe2x80x94[cycloalkyl-alkoxy)acetyl] mutilins; such as 14xe2x80x94Oxe2x80x94[(aminocycloalkyl-sulfanyl)acetyl]mutilins; 14xe2x80x94Oxe2x80x94[(aminocycloalkyl-alkyl-sulfanyl)acetyl] mutilins; 14xe2x80x94Oxe2x80x94[(aminocycloalkoxy)acetyl] mutilins; and 14xe2x80x94Oxe2x80x94[(aminocycloalkyl-alkoxy)acetyl] mutilins; preferably 14xe2x80x94Oxe2x80x94[(aminocycloalkyl-sulfanyl)acetyl]mutilins; e.g. cycloalkyl is preferably (C3-12)cycloalkyl; cycloalkoxy is preferably (C3-12)cycloalkoxy; alkyl is preferably (C1)alkyl; and alkoxy is preferably (C1-4)alkoxy.
In another aspect the present invention provides a compound of formula 
wherein
R is hydrogen;
R1 is hydrogen or a group of formula 
wherein
X is sulphur, oxygen or NR10, wherein R10, is hydrogen or alkyl; and
R9 is amino, alkyl, aryl or heterocyclyl; and, if X is oxygen, R9 is additionally hydrogen;
Y is sulphur or oxygen;
R2 is hydrogen or one or more substituents, e.g. including substituents such as conventional in organic, e.g. (pleuro)mutilin, chemistry;
R4 is hydrogen or alkyl;
R5 is hydrogen or alkyl;
R3 and R3xe2x80x2 are hydrogen, deuterium, or halogen;
R6, R7 and R8 are hydrogen or deuterium;
m is a number selected from 0 to 4;
n is a number selected from 0 to 10; and
p is a number selected from 0 to 10; with the proviso that n plus p are at least 1 and
preferably less than 13.
In formula I, preferably
R is hydrogen;
R1 is hydrogen or a group of the formula xe2x88x92C(=X)R9,
wherein X is oxygen; and
R9, is alkyl, e.g. (C1-8)alkyl, such as (C1-4)alkyl, unsubstituted or substituted alkyl, e.g. substituted by groups which are conventional in organic, e.g. pleuromutilin, chemistry, such as one or more amino; e.g., if R9 is alkyl substituted by amino, R9 is preferably the residue of an amino acid, e.g. valine, e.g. said residue includes that part of an amino acid which remains if the carboxylic group is split off;
Y is sulphur;
R2 is hydrogen;
R4, R5, R3, Rxe2x80x23, R6, R7, and R8 are hydrogen;
m is o;
n is 3 or4;
p is 0 or 1; and
p plus n is 3 or4.
In another aspect, the invention provides a compound of the formula Ip
wherein R1p is hydrogen or the residue of an amino acid; e.g. valyl; e.g. R1p is a group of formula xe2x80x94COxe2x80x94R9p wherein R9p is the residue of an amino acid which remains if the carboxylic group is split off.
In formula Ip a group xe2x80x94NHxe2x80x94R1p may be in any position of the cyclohexyl ring system and is preferably in position 2, 3, or 4. Amino acid in the meaning of Rap includes any amino acid, preferably valine; and R9p is preferably a group xe2x80x94CH(NH2)xe2x80x94CH(CH3)2. The amine group in said amino acid residue may be unprotected or protected, e.g. by appropriate amino acid protection groups, e.g. such as conventional, for example tert.butoxycarbonyl; and is preferably unprotected.
In another aspect, the present invention provides a compound which is:
14xe2x80x94Oxe2x80x94[(aminocyclohexan-2-yl-sulfanyl)acetyl]mutilin,
14xe2x80x94Oxe2x80x94[(aminocyclohexan-3-yl-sulfanyl)acetyl]mutilin,
14xe2x80x94Oxe2x80x94[(aminocyclohexan-4-yl-sulfanyl)acetyl]mutilin,
14xe2x80x94Oxe2x80x94[(N-valyl-aminocyclohexan-2-yl)sulfanyl)-acetyl]-mutilin,
14xe2x80x94Oxe2x80x94[(N-valyl-aminocyclohexan-3-yl)sulfanyl)-acetyl]-mutilin, or
14xe2x80x94Oxe2x80x94[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin; including
14xe2x80x94Oxe2x80x94[(N-(R)-valyl-(R)-aminocyclohexan-2(R)-yl)sulfanyl)-acetyl]-mutilin,
14xe2x80x94Oxe2x80x94[(N-(R)-valyl-(R)-aminocyclohexan-2(S)-yl)sulfanyl)-acetyl]-mutilin,
14xe2x80x94Oxe2x80x94[(N-(R)-valyl-(R)-aminocyclohexan-3(R)-yl)sulfanyl)-acetyl]-mutilin,
14xe2x80x94Oxe2x80x94[(N-(R)-valyl-(R)-aminocyclohexan-3(S)-yl)sulfanyl)-acetyl]-mutilin;
14xe2x80x94Oxe2x80x94[(N-(R)-valyl-cis-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin, and
14xe2x80x94Oxe2x80x94[(N-(R)-valyl-trans-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin, e.g., in free (base) form or in the form of a salt, such as a hydrochloride.
Preferred compounds include: 14xe2x80x94Oxe2x80x94[(aminocyclohexan-4-yl-sulfanyl)acetyl]mutilin and 14xe2x80x94Oxe2x80x94[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin. More preferred is 14xe2x80x94Oxe2x80x94[(N-valyl-aminocyclohexan-4-yl)sulfanyl)-acetyl]-mutilin.
A compound provided by the present invention is hereinafter designated as xe2x80x9cA compound of the present inventionxe2x80x9d. The present invention includes a compound of the present invention, e.g. including a compound of formulae I and Ip, in free (base) form and in the form of a salt, e.g. in the form of a solvate.
In another aspect, the present invention provides a compound of the present invention in the form of a salt, e.g. and in the form of a salt and in the form of a solvate, or in the form of a solvate.
A salt of a compound of the present invention includes a pharmaceutically acceptable salt, e.g. including a metal salt or an acid addition salt. Metal salts include for example alkali or earth alkali salts; acid addition salts include salts of a compound of the present invention with an acid, e.g. hydrogen fumaric acid, fumaric acid, naphthalin-1,5-sulfonic acid, hydrochloric acid, and deuterochloric acid; preferably hydrochloric acid or deuterochloric acid.
A compound of the present invention in free form may be converted into a corresponding compound in the form of a salt; and vice versa. A compound of the present invention in free form or in the form of a salt and in the form of a solvate may be converted into a corresponding compound in free form or in the form of a salt in unsolvated form; and vice versa.
A compound of the present invention may exist in the form of isomers and mixtures thereof; e.g. a compound of the present invention may contain asymmetric carbon atoms and may thus exist in the form of diastereoisomeres and mixtures thereof. For example, in a compound of formula Ip, wherein the group xe2x80x94NHxe2x80x94R1p is in position 2 or 3 of the cyclohexyl ring, the carbon atom of the cyclohexyl ring which is attached to the side chain of the mutilin ring and the carbon atom of the cyclohexyl ring to which the group xe2x80x94NHxe2x80x94R1p is attached are both asymmetric carbon atoms. A compound of formula Ip wherein the group xe2x80x94NHxe2x80x94R1p is in position 2 or 3 of the cyclohexyl ring may thus exist in (R) and (S) configurations in respect to both of these carbon atoms. For example, if R1p is the residue of an amino acid, that amino acid may comprise asymmetric carbon atoms. E.g., if R1p is valyl, the carbon atom to which the amine group of said valyl is attached is an asymmetric carbon atom. A compound of formula Ip wherein R1p is valyl may thus exist in (R) and in (S) configurations in respect to said valyl carbon atom.
Isomeric or diastereoisomeric mixtures may be separated as appropriate, e.g. according to conventional methods, to obtain pure isomers or diastereoismers, respectively. The present invention includes a compound of the present invention in any isomeric and diasteroisomeric form and in any isomeric and diastereoisomeric mixture. Preferably the configuration in the mutilin ring of a compound of formula I is the same as in a naturally produced pleuromutilin.
A compound of the present invention may be obtained as appropriate, e.g. according to, e.g. analogously, to any conventional method. E.g., 14-O-[(cycloalkyl-sulfanyl)acetyl]mutilins; 14xe2x80x94Oxe2x80x94[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins; 14xe2x80x94Oxe2x80x94[(cycloalkoxy)acetyl] mutilins; and 14xe2x80x94Oxe2x80x94[(cycloalkyl-alkoxy)acetyl] mutilins of the present invention may be prepared by reacting a 14xe2x80x94Oxe2x80x94[(mercapto)acetyl]mutilin or a 14xe2x80x94Oxe2x80x94[(hydroxy)acetyl] mutilin, respectively, with a hydroxycyclalkyl, or a hydroxyalkyl-cycloalkyl, respectively, in an activated form, e.g. in the form of an ester with a sulfonic acid, and isolating a compound of the present invention from the reaction mixture obtained.
Any compound of the present invention and any intermediate in the preparation of a compound of the present invention may be obtained as appropriate, e.g. according, such as analogously, to a conventional method, e.g. or as specified herein, including the examples. A compound of formula I or Ip may be obtained. e.g. according. e.g. analogously, to a process for the preparation of 14xe2x80x94Oxe2x80x94[(cycloalkyl-sulfanyl)acetyl]mutilins; 14xe2x80x94Oxe2x80x94[(cycloalkyl-alkyl-sulfanyl)acetyl] mutilins; 14xe2x80x94Oxe2x80x94[(cycloalkoxy)acetyl] mutilins; and14xe2x80x94Oxe2x80x94[(cycloalkyl-alkoxy)acetyl] mutilins.
In another aspect, the present invention provides a process for the production of a compound of formula I comprising the steps
a. reacting a compound of formula II 
wherein Y, R3, Rxe2x80x23, R4, and R5 are as defined above and R6, R7, and R8 are hydrogen, with a compound of formula III 
wherein R, R1, R2, m, n, and p are as defined above, in an activated form, e.g. in the form of an ester with 4-toulenesulfonic acid (tosylate), or of an ester with methylsulfonic acid (mesylate), to obtain a compound of formula I; wherein R6, R7 and R8 are hydrogen and Y, R1, R, R2, R3, Rxe2x80x23, R4, R6, m, n, and p are as defined above, and, if desired, b. introducing deuterium into a compound of formula I obtained in step a, to obtain a compound of formula I, wherein Y, R1, R, R2, R3, Rxe2x80x23, R4, R5, m, n, and p are as defined above and R6, R7, and R8 are deuterium.
A compound of formula II is known or may be obtained according, e.g. analogously, to a conventional method. A compound of formula III may be obtained as appropriate, e.g. according, e.g. analogously, to a conventional process. A compound of formula III may preferably be prepared according to the following process, e.g. or as described in the examples:
A dihydroxycycloalkyl or a (hydroxyalkyl)(hydroxy)cycloalkyl respectively, wherein R2, m, p, and n are as defined above; may be reacted in solvent which is inert under the reaction conditions with 4-toluenesulfonic or methanesulfonic acid anhydride to obtain a corresponding di-tosyl/mesyl-oxycycloalkyl, or (tosyl/mesyl)oxyalkyl)(tosyl/mesyloxy) cycloalkyl respectively; which di-tosyl/mesyl-compound is further reacted with sodium azide to obtain a corresponding (tosyl/mesyloxy)(azido)cycloalkyl, or a (tosyl/mesyloxyalkyl)(azido)cycloalkyl, respectively. In a (tosyl/mesyloxy)(azido)-compound obtained the azido group is reduced, e.g. catalytically hydrogenated, to obtain the corresponding (tosyl/mesyloxy)(amino)cycloalkyl, or (tosyl/mesyloxyalkyl)(amino)cycloalkyl, respectively; which is a compound of formula 111, wherein the hydroxy group is tosylated/mesylated; wherein R and R1 are hydrogen and wherein R2, m, p and, n are as defined above. If desired, the amine group obtained by reduction of the azido group may be reacted with R9xe2x80x94C(=X)OH, wherein R9 and X are as defined above, in an activated form, e.g. if X is oxygen R1xe2x80x94C(=X)OH may be in the form of an anhydride, halogenide; to obtain a compound of formula III, wherein R is hydrogen; R1 is a group of formula xe2x80x94C(=X)R9, wherein X and R9 are as defined above; and wherein R2, m, p, and n are as defined above.
Replacement of hydrogen atoms in a compound of formula 1, e.g. in the form of a salt; by deuterium atoms may be carried out as appropriate, e.g. according, e.g. analogously, to a conventional method, e.g. or according to a method described herein; e.g. by treatment of a compound of formula 1, e.g. including a compound of formula Ip; with deuterochloric acid (DCI) in appropriate solvent (system) and isolation of a compound of formula I, e.g. in the form of a salt, wherein hydrogen atoms, e.g. in the meaning of R6, R7, and R8 are replaced by deuterium atoms.
The production of a compound of formula I, wherein R3 and Rxe2x80x23 are deuterium or halogen may be carried out as appropriate, e.g. according, e.g. analogously, to a conventional method, e.g. via treatment of a compound of formula V 
wherein the carbon atoms carrying R3 and Rxe2x80x23, which both are hydrogen, together form a double bond and which is a known compound, with deuterium or with halogen, e.g. with F2, Cl2, or, Br2, to obtain a compound of formula V, wherein R3 and Rxe2x80x23 are deuterium or halogen; and further reacting a compound of formula V, wherein R3 and Rxe2x80x23 are deuterium or halogen as appropriate, e.g. according, e.g. analogously, to a conventional method, to obtain a compound of formula II, wherein, R3 and Rxe2x80x23 are deuterium or halogen and R6, R7, and R8 are hydrogen.
Preferably a compound of formula II may be obtained from a compound of formula V by reacting a compound of formula V with a compound of formula III 
wherein Y, R4, and R5 are as defined above and Hal is halogen, preferably bromo or chloro. A compound of formula III is known or may be obtained as appropriate, e.g. according, e.g. analogously, to a conventional method.
The compounds of the present invention, e.g. including a compound of formulae I and Ip, hereinafter designated as xe2x80x9cactive compound(s) of the present inventionxe2x80x9d exhibit pharmacological activity and are therefore useful as pharmaceuticals. For example, the active compounds of the present invention show antimicrobial, e.g. antibacterial, activity against gram positive bacteria, such as Staphylococci, e.g. Staphylococcus aureus, Streptococci, e.g. Streptococcus pyogenes, Streptococcus pneumoniae, Enterococci, e.g. Enterococcus faecium, as well as against mycoplasms, Chlamydia, and obligatory anaerobes, e.g. Bacteroides fragilis; in vitro in the Agar Dilution Test or Microdilution Test according to National Commitee for Clinical Laboratory Standards (NCCLS) 1997, Document M7-A4 Vol.17, No. 2: xe2x80x9cMethods for dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobicallyxe2x80x94Fourth Edition, Approved Standardxe2x80x9d; and in the Anaerobic Bacteria TEST according to National Committee for Clinical Laboratory Standards (NCCLS) VOL. 13, No. 26; M11-A4, Methods for Antimicrobal Susceptibility Testing of Anaerobic Bacteria; Approved Standard; Fourth Edition (1997). For example, the compound of example 1 shows in vitro in the above indicated Agar Dilution Test and/or Microdilution Test against bacterial strains as mentioned above MIC values of 0.01 to 1.0 xcexcg/mL.
In another aspect, the present invention provides a compound of the present invention for use as a pharmaceutical, preferably as an antimicrobial, such as an antibiotic, e.g. and an anti-anaerobic.
In a further aspect, the present invention provides a compound of the present invention for use in the preparation of a medicament for the treatment of microbial diseases, for example of diseases caused by bacteria, e.g. selected from Staphylococci, Streptococci, Enterococci; e.g. and of diseases caused by mycoplasms, Chlamydia, and obligatory anaerobes.
In a further aspect, the present invention provides a method of treatment of microbial diseases which comprises administering to a subject in need of such treatment an effective amount of a compound of the present invention e.g. in the form of a pharmaceutical composition.
For antimicrobial treatment, the appropriate dosage will, of course, vary depending upon, for example, the active compound of the present invention employed, the host, the mode of administration, and the nature and severity of the conditions being treated. However, in general, for satisfactory results in larger mammals, for example humans, an indicated daily dosage is in the range from about 0.5 to 3 g, of an active compound of the present invention conveniently administered, for example, in divided doses up to four times a day.
An active compound of the present invention may be administered by any conventional route, preferably orally, e.g. in form of tablets, powders, capsules, or suspensions; e.g. including non-resorbable oral formulations; or parenterally, e.g. in the form of injectable solutions or suspensions; or topically, e.g. in the form of nasal sprays, body solutions, creams, or eye drops. The active compounds of the present invention may be administered in analogous manner, e.g. in similar doses and for similar indications, as erythromycin(s), tetracycline(s). Surprisingly the active compounds of the present invention show also activity against strains which are resistant against erythromycin(s), tetracycline(s).
The active compounds of the present invention may be administered in the form of a pharmaceutically acceptable salt, e.g. an acid addition salt or metal salt; or in free form; optionally in the form of a solvate. The active compounds of the present invention in the form of a salt exhibit the same order of activity as the active compounds of the present invention in free form. The active compounds of the present invention may be administered in the form of pharmaceutical compositions.
In another aspect he present invention provides a pharmaceutical composition comprising a compound of the present invention, e.g. in free form or in the form of a pharmaceutically acceptable salt; e.g. and/or in the form of a solvate; in association with at least one pharmaceutical carrier or diluent.
Such compositions may be manufactured according, e.g. analogously, to a conventional method. Unit dosage forms may contain, for e.g., about 100 mg to about 1 g.
The active compounds of the present invention are additionally suitable as veterinary agents, e.g. veterinary active compounds, e.g. in the prophylaxis and in the treatment of microbial, e.g. bacterial diseases, in animals, such as fowl, pigs, and calves; e.g. and for diluting fluids for artificial insemination and for egg-dipping techniques.
In another aspect, the present invention provides a compound of the present invention for use as a veterinary agent.
In a further aspect, the present invention provides a compound of the present invention for the preparation of a veterinary composition which is useful as a veterinary agent.
The present invention further provides a veterinary method for the prophylaxis and in the treatment of microbial, e.g. bacterial diseases, which comprises administering to a subject in need of such treatment an effective amount of a compound of the present invention, e.g. in the form of a veterinary composition.
For use of the active compounds of the present invention as a veterinary agent, the dosage will of course vary depending upon the size and age of the animal and the effect desired. For example, for prophylactic treatment relatively low doses would be administered over a longer time period, e.g. 1 to 3 weeks. Preferred doses in drinking water are from 0.0125 to 0.05 weight by volume, particularly 0.0125 to 0.025; and in foodstuffs from 20 to 400 g/metric ton, preferably 20 to 200 g/metric ton. It is preferred to administer the active compounds of the present invention as a veterinary agent to hens in drinking water, to pigs in foodstuff, and to calves orally or parenterally, e.g. in the form of oral or parenteral preparations.